N-deacetylase/N-sulfotransferase 2 is an enzyme that catalyzes both N-deacetylation and N-sulfation (Wei Z, Swiedler S J, Ishihara M, Orellana A, Hirschberg C B. A single protein catalyzes both N-deacetylation and N-sulfation during the biosynthesis of heparan sulfate. Proc. Natl. Acad. Sci. USA. 90, 3885-3888. (1993); and U.S. Pat. No. 5,541,095). A transient expression system of human N-deacetylase/N-sulfotransferase 3 in monkey COS-7 cells has heretofore been used for the production of N-deacetylase/N-sulfotransferase (Aikawa J, Esko J D. Molecular cloning and expression of a third member of the heparan sulfate/heparin GlcNAc N-deacetylase/N-sulfotransferase family. J. Biol. Chem. 274, 2690-2695. (1999)), and however, was not suitable for obtaining the enzyme with stable quality in large amounts.
On the other hand, a method is known wherein a protein is extracellularly produced with a baculovirus system. For example, expression of Trypanosoma cruzi trans-sialidase in a baculovirus system using a gp67 signal sequence has been described in Marchal I, Cerutti M, Mir A M, Juliant S, Devauchelle G, Cacan R, Verbert A. Expression of a membrane-bound form of Trypanosoma cruzi trans-sialidase in baculovirus-infected insect cells: a potential tool for sialylation of glycoproteins produced in the baculovirus-insect cells system. Glycobiology. 11, 593-603 (2001). Expression of mouse TSH receptor in a baculovirus system using a gp67 signal sequence has been described in Vlase H, Matsuoka N, Graves P N, Magnusson R P, Davies T F. Folding-dependent binding of thyrotropin (TSH) and TSH receptor autoantibodies to the murine TSH receptor ectodomain. Endocrinology. 138, 1658-1666 (1997). Expression of single-chain antibodies in a baculovirus system using a gp67 signal sequence has been described in Kretzschmar T, Aoustin L, Zingel O, Marangi M, Vonach B, Towbin H, Geiser M. High-level expression in insect cells and purification of secreted monomeric single-chain Fv antibodies. J. Immunol. Methods. 195, 93-101 (1996). Expression of interleukin 1-related T1 receptor in a baculovirus system using a gp67 signal sequence has been described in Rupp B, Rossler U, Lowel M, Werenskiold A K. High level expression of the IL-1 receptor related T1 receptor in insect cells. Biochem. Biophys. Res. Commun. 216, 595-601 (1995). Moreover, expression of human AIDS virus envelop protein gp120 in a baculovirus system using a gp67 signal sequence has been described in Murphy C I, McIntire J R, Davis D R, Hodgdon H, Seals J R, Young E. Enhanced expression, secretion, and large-scale purification of recombinant HIV-1 gp120 in insect cell using the baculovirus egt and p67 signal peptides. Protein Expr. Purif. 4, 349-357. (1993). Erratum in: Protein Expr. Purif. 5, 103 (1994).
Moreover, polypeptide expression using a baculovirus-cultured insect cell system has been described in JP Patent Publication (Kokai) No. 2002-325579A (2002) and year Heisei 13, Riken research annual report, 681-687, Structural Biochemistry Laboratory, 4. Construction of Highly Efficient Protein Expression System Using Insect Cells, (1) Expression-Enhancing Factor (Sano and Maeda Y.; and Maeda K. (Cellular Signaling Laboratory)), wherein drastic increase in polypeptide expression level is seen by inserting a lobster-derived nontranslated leader sequence L21 upstream of the coding sequence of the polypeptide.
Furthermore, a case has been introduced recently wherein the amino acids of human N-deacetylase/N-sulfotransferase 2 are expressed in the form of a fusion protein with a His tag in a baculovirus system (Balagurunathan Kuberan, et al., Nature Biotechnology 1 Nov. 2003; 21, 1343-1346).